Medium supplement for high-yield industrial culture of fastidious anaerobes and medium composition containing the same

Abstract

A medium supplement for high-yield culture of anaerobes is disclosed. The medium supplement includes N-acetylhexosamine, L-aspartic acid, L-cysteine and cobalamin. A culture medium including the supplement and a culture method using the culture medium are also disclosed. It is possible to provide an innovative method which is capable of achieving high-concentration culture of anaerobes that are difficult to culture in high yield. The method is cost-effective, and in particular, is capable of culturing large amounts of fastidious aerobes suitable for use in food and pharmaceutical applications.

The invention claimed is: 
     
       1. A medium composition for culture of anaerobes, the medium composition comprising: plant peptone; yeast extract; potassium phosphate dibasic; as a carbon source, maltose and lactose; and as supplements, N-acetylhexosamine, an amino acid mixture of L-aspartic acid and L-cysteine, and cobalamin,
 wherein the anaerobes are selected from the group consisting of  Faecalibacterium prausnitzii, Anaerostipes caccae, Akkermansia muciniphila , and  Bifidobacterium longum.    
 
     
     
       2. The medium composition of  claim 1 , containing 20 g/L of plant peptone, 10 g/L of yeast extract, 2.5 g/L of potassium phosphate dibasic, 2.5 g/L of maltose, 2.5 g/L of lactose, 5 g/L of N-acetylhexosamine, 8 g/L of L-aspartic acid, 0.5 g/L of L-cysteine, and 0.0001 to 0.005 g/L of cobalamin. 
     
     
       3. The medium composition of  claim 1 , wherein the plant peptone is selected from the group consisting of soy peptone, wheat peptone, cotton peptone, pea peptone, broadbean peptone, lupin peptone, and potato peptone. 
     
     
       4. The medium of  claim 1 , wherein the anaerobes are  Faecalibacterium prausnitzii  or  Akkermansia muciniphila.    
     
     
       5. A method for culturing anaerobes, the method comprising inoculating anaerobes into the medium composition of  claim 1  and culturing the inoculated anaerobes under anaerobic conditions. 
     
     
       6. The method of  claim 5 , wherein the culturing is performed under conditions of a pH of 6.6 to 7.0, a culture temperature of 35 to 39° C., an agitation speed of 40 to 50 rpm, a nitrogen saturation degree of 80 to 90%, a hydrogen saturation degree of 0 to 5%, and a carbon dioxide saturation degree of 5 to 20%. 
     
     
       7. The method of  claim 5 , wherein the cultured anaerobes reach a cell density corresponding to a viable cell count of 10 10  CFU/mL or more as measured by a plate count method. 
     
     
       8. The method of  claim 5 , wherein a nitrogen saturation degree is 80%, a hydrogen saturation degree is 0%, and a carbon dioxide saturation degree is 20%. 
     
     
       9. The method of  claim 5 , wherein the anaerobes are  Faecalibacterium prausnitzii  or  Akkermansia muciniphila.    
     
     
       10. A method for culturing anaerobes, the method comprising inoculating anaerobes into the medium composition of  claim 2  and culturing the inoculated anaerobes under anaerobic conditions.